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Development of novel fluorescence-based and label-free noncanonical G4-quadruplex-like DNA biosensor for facile, specific, and ultrasensitive detection of fipronil

Fipronil is a broad-spectrum insecticide widely used in agriculture and residential areas; its indiscriminate use leads to environmental pollution and poses health hazards. Early detection of fipronil is critical to prevent the deleterious effects. However, current insecticide analysis methods such...

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Published in:Journal of hazardous materials 2022-04, Vol.427, p.127939-127939, Article 127939
Main Authors: Trinh, Kien Hong, Kadam, Ulhas Sopanrao, Rampogu, Shailima, Cho, Yuhan, Yang, Kyung-Ae, Kang, Chang Ho, Lee, Keun-Woo, Lee, Kyun Oh, Chung, Woo Sik, Hong, Jong Chan
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Language:English
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Summary:Fipronil is a broad-spectrum insecticide widely used in agriculture and residential areas; its indiscriminate use leads to environmental pollution and poses health hazards. Early detection of fipronil is critical to prevent the deleterious effects. However, current insecticide analysis methods such as HPLC, LC/MS, and GC/MS are incompetent; they are costly, immobile, time-consuming, laborious, and need skilled technicians. Hence, a sensitive, specific, and cheap biosensor are essential to containing the contamination. Here, we designed two novel biosensors—the first design relied on fluorescent labeling/quenching, while the second sensor focused on label-free detection using Thioflavin T displacement. Altogether, we identified four candidate aptamers, predicted secondary structures, and performed 3D molecular modeling to predict the binding pocket of fipronil in FiPA6B aptamer. Furthermore, the aptameric sensors showed high sensitivity to fipronil of sub-ppb level LOD, attributed to stringent experimental design. The biosensors displayed high specificity against other phenylpyrazole insecticides and demonstrated robust sensitivity for fipronil in real samples like cabbage and cucumber. Notably, to the best of our knowledge, this is the first demonstration of noncanonical G4-quadruplex-like aptamer binding to fipronil, verified using CD spectroscopy. Such aptasensors possess considerable potential for real-time measurements of hazardous insecticides as point-of-care technology. [Display omitted] •Ultrasensitive biosensors are crucial to detect and prevent toxic insecticide contamination.•Four unique ssDNA aptameric sensors are developed to detect fipronil.•Demonstrated two modes of detection: fluorescence labeling and label-free.•The aptamers displayed very high binding affinity (Kd ) and specificity.•Achieved detection sensitivity of the sub-ppb level.
ISSN:0304-3894
1873-3336
DOI:10.1016/j.jhazmat.2021.127939